Different gene expression profiles in iPSC-derived motor neurons from ALS8 patients with variable clinical courses suggest mitigating pathways for neurodegeneration.

Amyotrophic lateral sclerosis type 8 (ALS8) is an autosomal dominant form of ALS, which is caused by pathogenic variants in the VAPB gene. Here we investigated five ALS8 patients, classified as 'severe' and 'mild' from a gigantic Brazilian kindred, carrying the same VAPB mutation but displaying different clinical courses. Copy number variation and whole exome sequencing analyses in such individuals ruled out previously described genetic modifiers of pathogenicity. After deriving induced pluripotent stem cells (iPSCs) for each patient (N = 5) and controls (N = 3), motor neurons were differentiated, and high-throughput RNA-Seq gene expression measurements were performed. Functional cell death and oxidative metabolism assays were also carried out in patients' iPSC-derived motor neurons. The degree of cell death and mitochondrial oxidative metabolism were similar in iPSC-derived motor neurons from mild patients and controls and were distinct from those of severe patients. Similar findings were obtained when RNA-Seq from such cells was performed. Overall, 43 genes were upregulated and 66 downregulated in the two mild ALS8 patients when compared with severe ALS8 individuals and controls. Interestingly, significantly enriched pathways found among differentially expressed genes, such as protein translation and protein targeting to the endoplasmic reticulum (ER), are known to be associated with neurodegenerative processes. Taken together, the mitigating mechanisms here presented appear to maintain motor neuron survival by keeping translational activity and protein targeting to the ER in such cells. As ALS8 physiopathology has been associated with proteostasis mechanisms in ER-mitochondria contact sites, such differentially expressed genes appear to relate to the bypass of VAPB deficiency.

Different gene expression profiles in iPSC-derived motor neurons from ALS8 patients with variable clinical courses suggest mitigating pathways for neurodegeneration

Amyotrophic Lateral Sclerosis type 8 (ALS8) is an autosomal dominant form of ALS, which is caused by pathogenic variants in the VAPB gene. Here we investigated five ALS8 patients, classified as ‘severe’ and ‘mild’ from a gigantic Brazilian kindred, carrying the same VAPB mutation but displaying different clinical courses. Copy Number Variation (CNV) and Whole Exome Sequencing (WES) analyses in such individuals ruled out previously described genetic modifiers of pathogenicity. After deriving induced pluripotent stem cells (iPSCs) for each patient (N=5) and controls (N=3), motor neurons were differentiated, and high-throughput RNA-Seq gene expression measurements were performed. Functional cell death and oxidative metabolism assays were also carried out in patients’ iPSC-derived motor neurons. The degree of cell death and mitochondrial oxidative metabolism were similar in iPSC-derived motor neurons from mild patients and controls, and were distinct from those of severe patients. Similar findings were obtained when RNA-Seq from such cells was performed. Overall, 43 genes were upregulated and 66 downregulated in the two mild ALS8 patients when compared with severe ALS8 individuals and controls. Interestingly, significantly enriched pathways found among differentially expressed genes, such as protein translation and protein targeting to endoplasmic reticulum (ER), are known to be associated with neurodegenerative processes. Taken together, the mitigating mechanisms here presented appear to maintain motor neuron survival by keeping translational activity and protein targeting to ER in such cells. As ALS8 physiopathology has been associated with proteostasis mechanisms in ER–mitochondria contact sites, such differentially expressed genes appear to relate to the bypass of VAPB deficiency.

Different gene expression profiles in iPSC-derived motor neurons from ALS8 patients with variable clinical courses suggest mitigating pathways for neurodegeneration

Amyotrophic Lateral Sclerosis type 8 (ALS8) is an autosomal dominant form of ALS, which is caused by pathogenic variants in the VAPB gene. Here we investigated five ALS8 patients, classified as ‘severe’ and ‘mild’ from a gigantic Brazilian kindred, carrying the same VAPB mutation but displaying different clinical courses. Copy Number Variation (CNV) and Whole Exome Sequencing (WES) analyses in such individuals ruled out previously described genetic modifiers of pathogenicity. After deriving induced pluripotent stem cells (iPSCs) for each patient (N=5) and controls (N=3), motor neurons were differentiated, and high-throughput RNA-Seq gene expression measurements were performed. Functional cell death and oxidative metabolism assays were also carried out in patients’ iPSC-derived motor neurons. The degree of cell death and mitochondrial oxidative metabolism were similar in iPSC-derived motor neurons from mild patients and controls, and were distinct from those of severe patients. Similar findings were obtained when RNA-Seq from such cells was performed. Overall, 43 genes were upregulated and 66 downregulated in the two mild ALS8 patients when compared with severe ALS8 individuals and controls. Interestingly, significantly enriched pathways found among differentially expressed genes, such as protein translation and protein targeting to endoplasmic reticulum (ER), are known to be associated with neurodegenerative processes. Taken together, the mitigating mechanisms here presented appear to maintain motor neuron survival by keeping translational activity and protein targeting to ER in such cells. As ALS8 physiopathology has been associated with proteostasis mechanisms in ER–mitochondria contact sites, such differentially expressed genes appear to relate to the bypass of VAPB deficiency.

Desenvolvimento, confecção e avaliação dos efeitos e segurança de um protótipo de órtese de quadril utilizado na reabilitação funcional de paciente submetido a artroplastia de Girdlestone / Development, manufacturing and evaluation of effects and safety of a hip orthosis prototype used in the functional rehabilitation of a patient undergoing Girdlestone arthroplasty

A Artroplastia de Ressecção tipo Girdlestone (ARG) é um procedimento que altera o estilo de vida do indivíduo gerando fadiga precoce, instabilidade articular, distúrbio da marcha e discrepância de membros. Como medida auxiliar no processo de reabilitação podem ser utilizadas órteses. Contudo, não há no mercado nacional dispositivos adequados, sendo necessária a importação com custo elevado. Assim, este estudo desenvolveu e avaliou os efeitos e a segurança de um protótipo de órtese de quadril em uma paciente feminina submetida à ARG. O modelo proposto melhorou o desempenho físico e a funcionalidade e promoveu o alívio de dores osteomusculares. (AU)

Girdlestone resection arthroplasty (GRA) is a procedure that changes one's lifestyle, leading to early fatigue, joint instability, gait disorder and limb discrepancy. Orthoses can be used as an aid in the rehabilitation process. However, there are no adequate devices available in Brazil and the imported ones are high-cost. Thus, this study developed and evaluated the effects and safety of a hip orthosis prototype in a woman who underwent GRA. The proposed model improved physical performance and functionality and provided relief of musculoskeletal pain. (AU)

Anatomy and histochemistry of leaves and stems of Sapium glandulosum

Abstract Sapium belongs to Euphorbiaceae family and comprises 23 species. Sapium glandulosum (L.) Morong is popularly known in Brazil as "pau-leiteiro" and "leitosinha" and it is used in traditional medicine to cicatrisation. Its leaf extracts have shown analgesic, anti-inflammatory and antibacterial activities. The preliminary set of pharmacognostic tools used for quality assessment of medicinal plant parts is macro- and micro-anatomy and S. glandulosum has not anatomical and histochemical description. Thus the aim of this study was to investigate the anatomical and histochemical characteristics of the leaf and stem of S. glandulosum as a means of providing information for quality assessment of herbal industry. The leaves and stems were investigated by employing field emission scanning electron microscopy, light microscopy, and histochemistry techniques. The analysis showed that S. glandulosum had the following anatomical features: dorsiventral and amphistomatic leaves; paracytic stomata; tabular crystal druses; non-articulated and branched laticifers; midrib's biconvex shape with vascular systems in open arc with invaginated ends; petiole with a round shape and slight concavity on the adaxial side; six collateral vascular bundles in U-shaped organisation; a circular stem shape and a sclerenchymatous ring. In the histochemical tests lipophilic components were found in cuticle and in the latex; phenolic compounds were met in the mesophyll and in the latex; starch grains were found in the parenchymatous sheath; lignified elements were met in the sclerenchymatous ring in the cortex and in the perivascular sclerenchymatous caps, beyond in the vessel elements. These features are helpful when conducting a quality control process.

Euphorbia umbellata bark extracts – an in vitro cytotoxic study

ABSTRACT Euphorbia umbellata (Pax) Bruyns, Euphorbiaceae, is commonly used in folk medicine of southern Brazil to treat several kinds of cancer. The latex (part of the plant used for this purpose) is mixed with water and taken as treatment; but this matrix contains toxic potential related to the presence of some phorbol type diterpenes. So the aim of this study was to evaluate the cytotoxicity of the crude extract of the bark of E. umbellata and its fractions (Hex, CHCl3, EtOAc and MeOH) using in vitro assay (applying Jurkat cells line). A preliminary cytotoxic study (MTT reduction, trypan blue exclusion and DNA quantification assays) was executed to identify the most active material. The CHCl3 fraction displayed the highest activity and was selected for further investigation of any cytotoxic mechanism and evaluation of chemical composition; flow cytometry, Acridine orange and Hoechst 33342 staining experiments and Gas chromatography–mass spectrometry analysis were applied to achieve these results. This fraction demonstrated the best cytotoxic results against Jurkat cells line with IC50 of 29.00 ± 1.49, 10.06 ± 1.48 and 4.83 ± 2.25 µg/ml for 24, 48 and 72 h of experiment, respectively (trypan blue exclusion). The mechanism responsible for this action can be associated with the promotion of cell cycle arrest and apoptosis. The two main classes of compounds present in the CHCl3 fraction are steroids and triterpenes. Further, phytochemical studies with this fraction need to be evaluated, to try isolating these substances and establishing a more detailed cytotoxic study against Jurkat cells.

Cytotoxic biomonitored study of Euphorbia umbellata (Pax) Bruyns.

ETHNOPHARMACOLOGICAL RELEVANCE: Euphorbia umbellata latex (sap) has normally been used in folk medicine in southern Brazil to treat different types of cancers. AIM OF STUDY: To carry out a biomonitored investigation of partitioned latex using in vitro assay, to identify the main mechanisms related with the action of the most active fraction as well as to develop a phytochemical study with this material. MATERIALS AND METHODS: Biological screening was performed with hexane, chloroform, ethyl acetate and methanol fractions from the latex of E. umbellata using MTT, trypan blue, and neutral red assays to determine the cytotoxicity against HRT-18, HeLa and Jurkat cells and flow cytometry, DNA quantification, acridine orange and Hoechst 33342 staining to investigate mechanisms of action for the hexane extract. The phytochemical study of the hexane fraction was performed by chromatographic procedures and the substances were identified by NMR analysis. The isolated terpenes were evaluated using MTT to determine the cytotoxicity against Jurkat cells. RESULTS: All the fractions presented concentration and time dependent cytotoxicity. The hexane fraction showed the highest cytotoxicity; whereas the Jurkat cell was the lineage with the highest sensitivity (IC50 1.87µg/mL). Fragmentation of DNA and apoptosis are two mechanisms related with the toxicity of hexane fraction. The hexane fraction arrested the cell cycle in the G0/G1 phase, and the selectivity index was 4.30. Phytochemical study of the hexane fraction led to isolation of euphol (main compound) and germanicol acetate. Both substances demonstrated some slight cytotoxic activity against Jurkat cells after 72h; however the activity was minimal compared to vincristine (anticancer standard drug). CONCLUSION: The current research proves that the fractions of the latex from E. umbellata have a cytotoxic effect against three different cancer cells lines. The hexane fraction showed high in vitro cytotoxic effects against Jurkat cells demonstrating that the effect may be due to non-polar constituents. The two isolated terpenes (euphol and germanicol acetate) showed poor cytotoxic activity indicating that the anticancer properties of the extract may be caused by other substances present in the hexane fraction.

Cytotoxicity of latex and pharmacobotanical study of leaves and stem of Euphorbia umbellata (Janaúba)

AbstractIn southern Brazil, the bottled latex of Synadenium grantii Hook f., Euphorbiaceae, is popularly used as a treatment of all types of cancer. Similarly, Synadenium umbellatum Pax. is used in the central western region of Brazil for the same purpose and in the same manner of use. Both plants are popularly known as janaúba or leitosinha. The objectives of this study were to use pharmacobotanical analysis to verify whether these two species, which are considered to be distinct, are actually the same to determine anatomical markers; to assist in the identification and differentiation of other Euphorbia; and to evaluate the cytotoxic activity of the latex in relation to HeLa and HRT-18 cells. Leaves and stems of the species were collected in Goiânia and Ponta Grossa and were investigated using scanning electron microscopy and optical microscopy techniques. The latex was also collected and analyzed in relation to its cytotoxic effect by employing MTT and NR techniques. The pharmacobotanical study of the specimens in both localities showed that they were the same species, namely Euphorbia umbellata (Pax) Bruyns, which is the scientific nomenclature accepted and confirmed by an expert taxonomist who specializes in Euphorbia. The pharmacobotanical characteristics highlighted in this study can assist in the identification of the taxon and contribute to the control of the quality of this plant drug. The evaluation of the latex in relation to HRT-18 cells demonstrated action after 48 h of experiment. In contrast, in relation to HeLa cells its induced cytotoxicity in all times and a dose-dependent manner. The IC50 values (72 h) observed were 252.58 ± 18.51 µg/ml and 263.42 ± 15.92 µg/ml to MTT experiment and 250.18 ± 19.48 µg/ml and 430.56 ± 19.71 µg/ml to NR experiment for the HeLa and HRT-18 cells, respectively.

Análise da produção de sentidos em narrativas de afásicos participantes de grupo de convivência / Analyzing sense production in narratives for aphasic participants of the conviviality group

OBJETIVO: analisar os sentidos discursivos observados na fala de afásicos participantes de um grupo de convivência. MÉTODOS: o estudo foi realizado no Departamento de Fonoaudiologia da Universidade Federal de Pernambuco, onde foi formado o grupo de convivência de afásicos, constituído de seis sujeitos afásicos e cinco não-afásicos. Para o desenvolvimento das atividades do estudo, os encontros foram gravados e posteriormente transcritos, seguindo o sistema padronizado do NURC (Norma Universal de Transcrição), e em seguida realizada a análise. RESULTADOS: através das transcrições, foi possível encontrar os temas comuns e recorrentes ao discurso de afásicos, foram eles: incapacidades; lembranças e conhecimentos; memórias e épocas da vida; família; religião; trabalho e doenças/acidente vascular cerebral. Os temas de maior prevalência e iniciados pelos afásicos foram sobre as incapacidades, doenças e causas do acidente vascular cerebral, principalmente as alterações linguísticas. Verificou-se que o discurso desses sujeitos, independentemente das alterações que apresentaram, está impregnado de sentidos verdadeiros, que se produzem em decorrência da interação social entre os interlocutores, sujeitos ativos nessa troca de informações e constituídos de ideologia e história. CONCLUSÃO: pode-se observar que, mesmo possuindo um discurso reduzido, os afásicos apresentaram comportamentos e tentativas de interação com o outro, estabelecendo processos de significação.

PURPOSE: to analyze the discursive meanings observed in the speech of aphasic subjects in a coexistence group. METHODS: the study was conducted at the Speech Therapy department, from Universidade Federal de Pernambuco, where the aphasic coexistence group was formed, constituting by six aphasic subjects & five non - aphasic. For developing this study's activities, the discussions were recorded and later transcribed, according to the standardized system: NURC (Universal Standard of Transcription), and then we performed the analysis. RESULTS: through the transcripts, it was possible to find the common and the recurring themes of the aphasic speech. They were: disability; memories and knowledge, memories and life phases, family, religion, labor and disease/VEA (Vascular Encephalic Accident).The most common themes initiated by the aphasic subjects were about disabilities, diseases and VEA causes, especially about the linguistic changes. It was found that these subjects' speech, regardless of the changes and problems that they have or have had, is full of real meanings, which are produced as a result of the interaction between the social partners, who are active subjects, on this information exchange, and who are constructed by and ideology and a history. CONCLUSION: we can see that, even having a reduced or limited speech, the aphasic subjects had behaviors and attempts or efforts to interact with the others, building meaning processes.

Short- and long-term effects of a neonatal low-protein diet in rats on the morphology of the larynx.

OBJECTIVE: The aim of this study was to investigate the short- and long-term effects of a maternal low-protein diet during lactation on offspring laryngeal morphology. Our hypothesis was that a neonatal low-protein diet during the critical period of development alters micro- and macroscopic structures of the larynx in adult rats. METHODS: Male Wistar rats were assigned to a control (casein 17%, n=24) or low-protein (casein 8%, n=24) group according to their mother's diet during lactation. Body weight gain and growth rate were recorded throughout the experiment. The larynx was removed from offspring at days 22 and 60 of life. The anteroposterior and laterolateral lengths of the cartilages epiglottis, thyroid, and cricoid were measured by a digital caliper. The supraglottis, glottis, infraglottis, and vocal cords were stained by hematoxylin-eosin and their structures were analyzed by a Scion Image Beta 4.0.2 program. RESULTS: Pups from mothers fed a low-protein diet showed a lower body weight gain. The laterolateral and anteroposterior lengths of the larynx were shorter in undernourished offspring at 22 d old. There were no differences in the structure of the supraglottis, glottis, and infraglottis between groups except for keratinization in pups from undernourished mothers. The microstructure of the vocal cords was changed only at 60 d old. CONCLUSION: Macroscopic structures of the larynx are sensitive to short-term effects of a neonatal low-protein diet. Vocal cord development can be studied within the context of programming because their microscopic structures are sensitive to the long-term effects of a low-protein diet during lactation.